Mutagenicity, co-mutagenicity and anti-mutagenicity of glyco-ursodeoxycholic acid (GUDCA) were examined by the Ames assay using Salmonella typhimurium strain TA98 with S9. As pro-mutagens, 2-aminoanthracene (2AA), Benzo[a]pyrene (BaP), 3-amino-1-dimethyl-5H-pyrido[4, 3-b]indole (Trp-P-2), 2-amino-3-methylimidazo[4, 5-f]quinoline (IQ) and 2-amino-3, 4-dimethylimidazo[4, 5-f]quinoline (MeIQ) were used. In addition to these pro-mutagens, blue-chitin extracts of human gallbladder bile (BCE) collected from the cholecystectomized patients with cholelithiasis were used in order to investigate the role of GUDCA on mutagen(s) actually existing in human bile. It was found that GUDCA did not show mutagenicity in this test system. Concerning the modification of mutagenic activities of pro-mutagens, GUDCA showed the different directions. GUDCA acted as co-mutagen, since it enhanced the mutagenic activities of 2AA and BaP. But, acted as anti-mutagen, since it suppressed the activities of Trp-P-2, IQ and MeIQ, all of which were classified as heterocyclic amines. GUDCA also suppressed the mutagen(s) in human bile. Because of the use of blue-chitin absorbed method for testing bile mutagenicity, the chemicals involved were considered to be heterocyclic amines and other polycyclic compounds. In these we suspect the bile mutagens are heterocyclic amines. Further examination should be directed towards the investigation into the mechanism of anti-mutagenic effects of GUDCA on mutagen(s) actually existing in human bile.
To examine the association of body mass index (BMI) and waist-to-hip ratio (WHR) with glucose intolerance among adults age 45 and over, we conducted a population-based study using an oral glucose tolerance test as a primary examination in two areas of Funagata, Japan, in 1990 and 1992. The number of eligible subjects was 1673. The participation rate was 84% (1408/1673). Glucose tolerance was assessed by the 1985 World Health Organization criteria as having diabetes (DM), impaired glucose tolerance (IGT), or normal glucose tolerance (NGT). Analyses by the generalized linear model revealed that both BMI and WHR were higher in the subjects with DM and IGT than those with NGT in both men and women after controlling for age. Analyses employing multiple logistic regression indicated that BMI and WHR were independently associated with IGT and DM in both men and women, except for BMI with IGT for men. The odds ratios for IGT associated with BMI were 1.06 (p=0.19) for men and 1.11 (p<0.01) for women. Those associated with WHR were 2.14 (p<0.01) for men and 1.35 (p<0.01) for women. These results imply that WHR plays an important role for developing DM independent of BMI.
We have newly found that interleukin-2 (IL-2) increases mouse voluntary running 24 hours, but not 30 minutes, after the injection. We suspected that IL-2 induced a substance increasing the voluntary running for 24 hours after injection. Serum obtained from mice 24 hours after the IL-2 treatment was fractioned with the use of an ion-exchanger and an ultra-filtration method, and the amino acid sequence analysis indicated that the substance purified from the effective fraction was a fragment of mouse complement 3a (C3a) lacking the primary 9 amino acids. The 20 amino acid peptide synthesized according to the fragment showed the activity increasing the voluntary running, but the 20 amino acid peptide synthesized according to the C3a itself did not. The effect of the synthesized peptide was demuted by haloperidol but not by a specific dopamine 2 antagonist (−)sulpiride. The present findings clearly indicate that IL-2 produces the C3a fragment lacking the primary 9 amino acids which directly promotes the voluntary running, and that the effect of the fragment is mediated by an activity of haloperidol on the neurons, except for the dopamine 2 antagonism.
The aim of this study was to find vascular areas where clonidine decreases the regional vascular resistance when this drug lowers arterial pressure in conscious spontaneously hypertensive rats and normotensive control rats. Arterial pressure was observed with an indwelling catheter at a carotid. Blood flow was measured with an electromagnetic flow probe implanted around the renal artery or the superior mesenteric artery. Regional vascular resistance was calculated as arterial pressure divided by blood flow. Intravenous bolus injection of clonidine at a dose to decrease arterial pressure decreased renal resistance and superior mesenteric resistance. Quantitatively, the combined effect of the decrease in these two resistances was enough to account for the decrease in arterial pressure. Although clonidine is thought to inhibit sympathetic nerve activity centrally, the above vasodilator effect is not ascribable to this inhibitory mechanism: Sympathetic activity to be inhibited does not seem to be present in the superior mesenteric area and clonidine similarly decreased renal vascular resistance even after renal denervation.
Neuron-specific enolase (NSE) and carcinoembryonic antigen (CEA) levels in the culture supernatant of the 65 pulmonary carcinoma cell lines: Small cell lung cancer (SCLC) 18, large cell carcinoma 14, squamous cell carcinoma 14, adenocarcinoma 14 and adenosquamous cell carcinoma 5, were measured by a radioimmunoassay (RIA). The mean value of NSE was 30.8±22.4 ng/ml and 9.2±8.7 ng/ml in SCLC and non-SCLC, respectively. The mean value of CEA was 15.1±20.9 ng/ml and 26.6±72.3 ng/ml in SCLC and non-SCLC, respectively. A significant difference in NSE levels was obtained between SCLC cell lines and non-SCLC cell lines. In SCLC cell lines, a significant inverse proportional correlation was observed between NSE and CEA levels. The CEA production tended to be higher in cells with low levels of NSE than in those with high NSE production. With respect to correlation between marker production and growth characteristics of SCLC cells in vitro, significantly higher NSE and lower CEA levels were found in cells growing with floating colony or neurite like characteristics (classic cell type) than those in cells with epithelial or intermediate growth characteristics (variant cell type). A significant positive correlation between NSE levels and the survival periods was found in follow-up studies of 10 patients who underwent surgery with complete resection of the primary tumor. All of 4 long term survivors over 3 years after surgery had significantly high NSE and relatively low CEA producing tumors. The relationship of these markers to clinical status of the patient suggests that an analysis for correlation of NSE and CEA levels in SCLC patients may be useful to discriminate between a pure neuroendocrine SCLC tumor and a mixed small cell/large cell tumor, and in monitoring therapeutic effect and prognosis of each patient.
In human promyelocytic leukemia cell line HL60, apoptosis was induced by treatment with gossypol that is an inhibitor of protein kinase C. Gossypol acetic acid was added to HL 60 cells at 50, 100, 150 and 200 μM concentrations for six hours. Morphological features of apoptosis as well as internucleosomal DNA fragmentation were evaluated by light microscope, agarose gel electrophoresis and spectrofluorometric quantitation. Our results indicated that with the effective concentrations of gossypol (50 and 100 μM), apoptosis was induced in HL 60 cells.
Some normal people are falsely classified as having impaired glucose tolerance (IGT) if they are given an oral glucose tolerance test (OGTT) when their last meal contained very few carbohydrates. In this study, the duration of carbohydrate restriction was extended to one and three days and the relationship between the carbohydrate restriction and the glucose tolerance after an OGTT was examined. Two different groups of normal subjects were placed on high-carbohydrate (80% carbohydrates) and low-carbohydrate (10%) diets before an OGTT; one group for one day and the other for 3 days. None of the subjects showed impairment of glucose tolerance when placed on the high-carbohydrate regimens. In contrast, 3 of 12 subjects and 2 of 8 subjects placed on the low-carbohydrate diets for 1 and 3 days, respectively, were classified as having IGT. The impairment of glucose tolerance was invariably accompanied by an increase in the fasting plasma free fatty acid level. The longer the period of carbohydrate restriction, the severer was the glucose tolerance impairment. However, the number of subjects who were classified as having IGT did not depend on the duration of carbohydrate restriction. The impairment of glucose tolerance after carbohydrate restriction may be associated with the Randle effect, which is the activation of the glucose-free fatty acid cycle.
Granulocyte colony-stimulating factor (G-CSF) is a cytokine that regulates the proliferation, differentiation and survival of cells in the granulocytic lineage. In this study, however, we found that G-CSF or interleukin-6 (IL-6) induced UF-1, a human acute promyelocytic leukemia cell line, into apoptosis that was confirmed by morphological features and DNA fragmentation. This rare response is demonstrated for the first time with human acute promyelocytic leukemia cell line. The apoptosis induced by G-CSF or IL-6 was not preceded by terminal differentiation characterized by morphological maturation, capability to reduce nitroblue tetrazolium, or surface CD11b expression. Interestingly, Western blot analysis revealed that the stimulation of UF-1 with either G-CSF or IL-6 resulted in excessive activation of both signal transducer and activator of transcription 3α (Stat3α) and Stat3β. Furthermore, an additional 18 kDa Bax-related protein was expressed by the stimulation of G-CSF or IL-6, while Bcl-2 and Bcl-X proteins remained unchanged. These findings suggest that UF-1 may be a valuable tool in investigating the aberrant regulation of apoptosis, especially the Stat3 involvement in the mechanism of apoptosis induction.
Mouse forced swimming is one of the behavioral depression models and repeated electroconvulsive shock (ECS) treatment has been used to human depression. In order to investigate the mechanism of the anti-depressive effect induced by repeated ECS, we investigated the effect of repeated ECS with the mouse forced swimming model. The 5 times par-day ECS remarkably increased the typical anti-depressive behavior climbing 24 hours after the final treatment. The anti-depressive activity was declined by a dopamine 1 antagonist SCH-23390 at the doses of 1 and 0.1 mg/kg, but not by the other dopamine, serotonin and adrenoceptor antagonists at the dose of 1 mg/kg. The present findings strongly suggest that the late anti-depressive effect of repeated ECS is mediated by the dopamine 1 receptor activity. The present findings will also contribute to the further investigations of the effect of repeat ECS treatments on human depression.